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TitleProgrammable genome editing in human cells using RNA-guided bridge recombinases.
Journal, issue, pagesScience, Page eadz1884, Year 2026
Publish dateFeb 5, 2026
AuthorsOana Pelea / András Tálas / Javier Fernández Carrera / Nicolas Mathis / Lilly van de Venn / Charles D Yeh / Péter I Kulcsár / Kim F Marquart / Yanik Weber / Saskia E Gerecke / Isabelle F Harvey-Seutcheu / Dominic Mailänder / Moritz M Pfleiderer / Christelle Chanez / Jacob E Corn / Gerald Schwank / Martin Jinek /
PubMed AbstractSite-specific insertion of gene-sized DNA fragments remains an unmet need in the genome editing field. IS110-family serine recombinases have recently been shown to mediate programmable DNA ...Site-specific insertion of gene-sized DNA fragments remains an unmet need in the genome editing field. IS110-family serine recombinases have recently been shown to mediate programmable DNA recombination in bacteria using a bispecific RNA guide (bridge RNA) that simultaneously recognizes target and donor sites. Here, we show that the bridge recombinase ISCro4 is highly active in human cells, and provide structural insights into its enhanced activity. Using plasmid- or all-RNA-based delivery, ISCro4 supports programmable multi-kilobase exisions and inversions, and facilitates donor DNA insertion at genomic sites with efficiencies exceeding 6%. Finally, we assess ISCro4 specificity and off-target activity. These results establish a framework for the development of bridge recombinases as next-generation tools for editing modalities that are beyond the capabilities of current technologies.
External linksScience / PubMed:41642947
MethodsEM (single particle)
Resolution2.81 Å
Structure data

EMDB-55572, PDB-9t56:
Cryo-EM structure of ISCro4-DBL-TBL-tDNA-dDNA synaptic complex
Method: EM (single particle) / Resolution: 2.81 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • citrobacter rodentium icc168 (bacteria)
KeywordsRECOMBINATION / Nucleic acid interaction / RNA binding / DNA binding / tetramer / recombinase

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